A PRECEDE-PROCEED model-based educational intervention to promote healthy eating habits in middle school girls.

The present study was designed to develop Nutrition Education Program (NEP) based on PRECEDE-PROCEED model (PPM) to address healthy eating behavior among middle school girls aged between 4 and 12 years. For this, middle school girls from grade 1 to 8 (n = 900) were consulted for their eating behaviors, followed by the analysis of their health problems. From 15 different schools of three large cities (Faisalabad, Lahore, and Rawalpindi) of Pakistan, students were divided into two groups: control group (n = 30) and intervention group (n = 30) from each school. The data were collected through interview-based questionnaires according to the phases of PRECEDE Model and evaluated based on PROCEED model. Implementation of NEP was carried out through lectures. Lessons were prepared to enhance student's awareness about nutritious food and healthy lifestyle through educational pamphlets and influenced their attitude towards selection of food choices from My-Plate. Results showed that NEP was quite successful for long-term results. A significant increase in total caloric intake was observed after 8 weeks of NEP intervention (1694 ± 217 Kcal) as compared to before intervention (1329 ± 318 Kcal). Similarly, carbohydrate, protein, and fat content was also increased in daily diet. Conclusively, NEP based on PPM has great impact on healthy lifestyle of middle school girls. Significant difference was observed in score of health variables before and after NEP intervention.

Status of Intestinal Helminth Infection in Schools Implementing the Home-Grown School Feeding Program and the Impact of the Program on Pupils in Anambra State, Nigeria.

PURPOSE: Intestinal helminthiasis among pupils is a major impediment to their well-being and development. This 2-year study determined how home-grown school feeding programme (HGSFP) impacted on pupils' school attendance and created opportunity for more children to assess interventions geared towards the prevention and possible elimination of intestinal helminthiasis. METHODS: Faecal samples were collected from 1677 consented pupils and examined for helminths using Kato-Katz technique. Pupils were treated with 40 mg Levamisole (anti-helminthic) and re-examined. Structured questionnaires and focus group discussions were used to determine anthropometric indices and risk factors. RESULTS: A 6% prevalence rate which was not significant with respect to sex or age was observed. Helminth parasites observed were Ascaris lumbricoides (4.4%), Trichuris trichiura (0.8%), Hookworm spp. (0.2%), and Taenia spp. (0.5%). Two pupils (1.98%) recorded high intensity of infection (epg) while 98.02% had light infection. Ascaris lumbricoides voided following treatment were 290. The Body Mass Index (BMI) indicated that infected pupils had lower values (14.6317) compared to uninfected pupils (19.2003). The HGSFP significantly improved school attendance by 30%. Lack of knowledge about transmission (P = 0.0004), hand hygiene (P = 0.01) and usage of toilet (P = 0.01) were risk factors observed. CONCLUSION/SIGNIFICANCE: Intestinal helminthiasis remains a public health problem among pupils because environmental and behavioural risk factors persist. HGSFP improved school attendance and created opportunity for more inclusive participation of school-aged children for deworming. This has great potential in fast-tracking the elimination of intestinal helminthiasis. The present study consequently advocates the need to sustain HGSFP in schools.

Molecular diagnosis for the novel coronavirus SARS-CoV-2: lessons learnt from the Ghana experience.

BACKGROUND: A novel coronavirus, SARS-CoV-2 is currently causing a worldwide pandemic. The first cases of SARS-CoV-2 infection were recorded in Ghana on March 12, 2020. Since then, the country has been combatting countrywide community spread. This report describes how the Virology Department, Noguchi Memorial Institute for Medical Research (NMIMR) is supporting the Ghana Health Service (GHS) to diagnose infections with this virus in Ghana. METHODS: The National Influenza Centre (NIC) in the Virology Department of the NMIMR, adopted real-time Polymerase Chain Reaction (rRT-PCR) assays for the diagnosis of the SARS-CoV-2 in January 2020. Samples from suspected cases and contact tracing across Ghana were received and processed for SARS-CoV-2. Samples were 'pooled' to enable simultaneous batch testing of samples without reduced sensitivity. OUTCOMES: From February 3 to August 21, the NMIMR processed 283 946 (10%) samples. Highest number of cases were reported in June when the GHS embarked on targeted contact tracing which led to an increase in number of samples processed daily, peaking at over 7,000 samples daily. There were several issues to overcome including rapid consumption of reagents and consumables. Testing however continued successfully due to revised procedures, additional equipment and improved pipeline of laboratory supplies. Test results are now provided within 24 to 48 hours of sample submission enabling more effective response and containment. CONCLUSION: Following the identification of the first cases of SARS-CoV-2infection by the NMIMR, the Institute has trained other centres and supported the ramping up of molecular testing capacity in Ghana. This provides a blueprint to enable Ghana to mitigate further epidemics and pandemics. FUNDING: The laboratory work was supported with materials from the Ghana Health Service Ministry of Health, the US Naval Medical Research Unit #3, the World Health Organization, the Jack Ma Foundation and the University of Ghana Noguchi Memorial Institute for Medical Research. Other research projects hosted by the Noguchi Memorial Institute for Medical Research contributed reagents and laboratory consumables. The funders had no role in the preparation of this manuscript.

Interaction of linear homologous DNA duplexes via Holliday junction formation.

Interaction of linear homologous DNA duplexes by formation of Holliday junctions was revealed by electrophoresis and confirmed by electron microscopy. The phenomenon was demonstrated using a model of five purified PCR products of different size and sequence. The double-stranded structure of interacting DNA fragments was confirmed using several consecutive purifications, S1-nuclease analysis, and electron microscopy. Formation of Holliday junctions depends on DNA concentration. A thermodynamic equilibrium between duplexes and Holliday junctions was shown. We propose that homologous duplex interaction is initiated by nucleation of several dissociated terminal base pairs of two fragments. This process is followed by branch migration creating a population of Holliday junctions with the branch point at different sites. Finally, Holliday junctions are resolved via branch migration to new or previously existing duplexes. The phenomenon is a new property of DNA. This type of DNA-DNA interaction may contribute to the process of Holliday junction formation in vivo controlled by DNA conformation and DNA-protein interactions. It is of practical significance for optimization of different PCR-based methods of gene analysis, especially those involving heteroduplex formation.

Orientation tuning of shape from shading.

Four experiments were performed to assess the effect of different orientations and direction of lighting on the visual processing of shaded or bipartite disks. In the first two experiments, observers were presented with nine different shading orientations from 0 degree to 180 degrees. Targets were detected in a rapid and parallel fashion for shaded disks when the orientation of the shading gradient was not horizontal (90 degrees) or oriented at 67.5 degrees. Search asymmetries favoring the detection of "pock" targets over "ball" targets were found for all orientations. The search rates for bipartite disks were similar to the shaded disks at 0 degree, 22.5 degrees, and 90 degrees but not for intermediate orientations, and no search asymmetries were found. These differences suggest that shaded displays and bipartite displays are processed by different underlying mechanisms. The third experiment showed that the direction of the light source (left or right) had no influence on search asymmetries around the 90 degrees point. Shading gradient orientation affected magnitude estimates of depth in the fourth experiment. These experiments show that the visual system's "assumption" of overhead lighting is broadly tuned.

Correct heteroduplex formation for mutation detection analysis.

BACKGROUND: The majority of mutation detection methods for unknown mutations are polymerase chain reaction (PCR)-based methods dependent on the formation of heteroduplexes between wild-type and mutant strands of DNA. METHODS AND RESULTS: This report discusses the difficulties associated with forming heteroduplexes with a large DNA fragment and the implications for subsequent mutation detection by the chemical cleavage of mismatch technique and other methods reliant on heteroduplex formation. It was found that the size and sequence context of the fragment being investigated inhibited correct heteroduplex formation. The problem was overcome by dividing the sequence into two overlapping fragments. CONCLUSIONS: Early identification of this problem in other fragments will help with the rapid optimization of PCR-based mutation detection methods.

Investigation of a unique male and female sibship with Kallmann's syndrome and 46,XX gonadal dysgenesis with short stature.

A sibship is described where the brother and a sister both have Kallmann's syndrome (anosmia and deficiency of gonadotrophin releasing hormone) and the woman also has streak ovaries. Although there are several conditions that may occur with Kallmann's syndrome, there are no known reports of ovarian dysgenesis being associated with this disorder. Cytogenetic analysis showed no rearrangement or major deletions of the chromosomes. Linkage analysis using informative microsatellite markers predicts that a gene other than KAL1 (at Xp22.3) is implicated in the Kallmann's syndrome manifesting concurrently with ovarian dysgenesis found in this family.

Reactivity of potassium permanganate and tetraethylammonium chloride with mismatched bases and a simple mutation detection protocol.

Many mutation detection techniques rely upon recognition of mismatched base pairs in DNA hetero-duplexes. Potassium permanganate in combination with tetraethylammonium chloride (TEAC) is capable of chemically modifying mismatched thymidine residues. The DNA strand can then be cleaved at that point by treatment with piperidine. The reactivity of potassium permanganate (KMnO4) in TEAC toward mismatches was investigated in 29 different mutations, representing 58 mismatched base pairs and 116 mismatched bases. All mismatched thymidine residues were modified by KMnO4/TEAC with the majority of these showing strong reactivity. KMnO4/TEAC was also able to modify many mismatched guanosine and cytidine residues, as well as matched guanosine, cytidine and thymidine residues adjacent to, or nearby, mismatched base pairs. Previous techniques using osmium tetroxide (OsO4) to modify mismatched thymidine residues have been limited by the apparent lack of reactivity of a third of all T/G mismatches. KMnO4/TEAC showed no such phenomenon. In this series, all 29 mutations were detected by KMnO4/TEAC treatment. The latest development of the Single Tube Chemical Cleavage of Mismatch Method detects both thymidine and cytidine mismatches by KMnO4/TEAC and hydroxylamine (NH2OH) in a single tube without a clean-up step in between the two reactions. This technique saves time and material without disrupting the sensitivity and efficiency of either reaction.

Chemical cleavage of mismatch: a new look at an established method.

Chemical cleavage of mismatch (CCM), also known as chemical mismatch cleavage (CMC) or the HOT (hydroxylamine/osmium tetroxide) chemical method, has been used for detection of sequence variability with many systems since it was first described. Recently, adaptation to fluorescence-based detection systems has fundamentally changed both the execution and analysis of CCM. This review will outline major advances in the methodology of CCM, from the advent of PCR through fluorescent analysis, and includes applications and modifications of CCM.

A retrospective cost-effectiveness analysis of the treatment of onychomycosis in general practice.

Analysis of the computer records of 100 general practices from the CompuFile Doctors Independent Network revealed 1492 patients receiving treatment for onychomycosis in the first 6 months of 1994 with terbinafine, tioconazole, amorolfine or griseofulvin. These records indicated the average treatment time for each agent, number of general practitioner consultations and incidence of hospital referrals and minor surgery. Applying standard costs to this resource consumption gave the direct costs for each of these four agents. Published clinical and mycological cure rates allowed a cost per success to be calculated. These were as follows: terbinafine (n = 511) pound258, amorolfine (n = 315) pound312, griseofulvin (n = 196) pound356 and tioconazole (n = 470) pound520. Sensitivity analysis showed that terbinafine remained the most cost-effective option despite variations in resource costs. The cost impact on a typical practice for switching from a less to a more cost-effective treatment is discussed.

A genomewide linkage study of preeclampsia/eclampsia reveals evidence for a candidate region on 4q.

Preeclampsia (PE) and eclampsia (E) are potentially life-threatening conditions that can occur during human pregnancy. Generally considered to be different degrees of severity of the same disease process, the PE/E syndrome is thought to be predominantly genetic in origin, although its exact etiology and genetics are not fully understood. Here we report results of a genomewide linkage search for the gene(s) responsible for susceptibility to PE/E, using 15 informative pedigrees and 90 polymorphic DNA markers from all autosomes. Because of uncertainties concerning inheritance and diagnosis, four different models that assume maternal gene expression have been used to carry out LOD-score analysis. The region between D4S450 and D4S610 (2.8 cM) on the long arm of chromosome 4 was identified as a strong candidate region for a PE/E-susceptibility locus. The maximum multipoint LOD score within this interval was 2.9. Analysis of markers in the region around D4S450 and D4S610 by the affected-pedigree-member method also supported the possibility of a susceptibility locus in this region. However, to verify or exclude definitively linkage to this region, other groups of PE/E pedigrees will be required.

New MspI polymorphism at +83 bp of the human apolipoprotein AI gene: association with increased circulating high density lipoprotein cholesterol levels.

We recently showed that loss of a MspI restriction site in the 5'-end (intron 1) of the apolipoprotein (apo) AI gene is due to a C to T transition (+83 bp) and/or a G to A transition (+84 bp). Since this region may be relevant to the regulation of apo AI gene expression and therefore to plasma high density lipoprotein cholesterol (HDL-C), we explored the association between this MspI polymorphic site and circulating HDL-C levels in 243 healthy Caucasians. There were 143 aged 18-67 years (60 males and 83 females) and 100 aged 6-12 years (58 males and 42 females). We also compared this association with a known MspI polymorphic site, a G to A transition at -75 bp of the apo AI gene. The rare allele (-) frequency for the polymorphism at +83 bp was 4.1% and 22.1% for the polymorphism at -75 bp. Subjects heterozygous for the loss of the MspI restriction site at +83 bp (genotype: M2+-, n = 20) had higher HDL-C levels than M2+2 subjects (mean +/- SD: 1.73 +/- 0.31 vs. 1.41 +/- 0.39 mmol/l, P < 0.05 for adults; 1.71 +/- 0.33 vs. 1.34 +/- 0.29 mmol/l, P < 0.01 for children). Adults with the G to A substitution at -75 bp also had higher HDL-C levels (1.56 +/- 0.36 mmol/l for AA, 1.53 +/- 0.38 mmol/l for GA, and 1.36 +/- 0.38 mmol/l for GG, P < 0.05); this difference was not observed in the children. The MspI polymorphisms at both sites were in linkage disequilibrium. Their joint effect on the HDL-C levels was also significant and individuals with rare alleles (-) at both sites had the highest HDL-C levels. In an analysis of variance, the MspI polymorphism at +83 bp, and at -75 bp and gender independently accounted for 6.5%, 1.7%, and 5.9%, respectively, of the variance in circulating HDL-C levels when age was controlled as a covariate. We conclude that loss of the MspI site by the C to T (+83 bp) and/or the G to A (+84 bp) transitions is highly associated with increased HDL-C levels. The association appears to be more significant than that of the G to A transition at -75 bp.

HLA-G deletion polymorphism and pre-eclampsia/eclampsia.

OBJECTIVE: To investigate a HLA-G deletion polymorphism in pre-eclamptic pedigrees and the general population. DESIGN: A population association study of HLA-G genotypes from pre-eclamptic/eclamptic patients and control groups. SETTING: Analyses undertaken in the School of Biological Sciences, Macquarie University. Patients were from Royal Women's Hospital, Melbourne, and controls were from Westmead Hospital and Macquarie University, Sydney. SUBJECTS: One hundred and ninety-six individuals, consisting of 29 pre-eclamptic/eclamptic (PE/E) patients, 13 individuals born of a PE/E pregnancy, 46 blood relatives of PE/E patients, 21 husbands of PE/E patients, 25 women normotensive in first pregnancy, 15 husbands of women normotensive in first pregnancy and 47 staff and students of Macquarie University. RESULTS: Genotypic and gene frequencies were not significantly different in the seven groups examined. CONCLUSION: There is no detectable relationship between susceptibility to pre-eclampsia or being born of a pre-eclamptic pregnancy and HLA-G genotype.

C to T and/or G to A transitions are responsible for loss of a MspI restriction site at the 5'-end of the human apolipoprotein AI gene.

We detected the loss of a MspI restriction site by a C to T transition at +83 bp and a G to A transition at +84 bp of the 5'-end non-coding region of the human apolipoprotein AI gene. This base change occurred at the "hot spot" (CCGG) for methylation, which may be important in the regulation of gene expression. The population frequency for the loss of the MspI site is 6.1%.